ANKEF1 Knockout HEK293T Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal cell population in which the ANKEF1 gene has been disrupted to generate a loss-of-function model. This gene-edited product is derived from HEK293T cells and is supplied as a polyclonal pool, representing a heterogeneous mixture of edited alleles suitable for studying ANKEF1-dependent cellular functions without the selective pressure of single-cell cloning. The use of CRISPR/Cas9-mediated gene disruption enables robust suppression of target protein expression, providing a powerful tool for functional genomics and pathway analysis in a widely used human cell line.
The HEK293T cell line, a derivative of the human embryonic kidney 293 cell line, is immortalized with adenovirus type 5 DNA and constitutively expresses the SV40 large T antigen. This genetic background supports high transfection efficiency and episomal replication of plasmids containing the SV40 origin, making HEK293T cells a standard host for recombinant protein production and lentiviral packaging. Their epithelial origin and renal lineage provide a relevant context for studying calcium signaling and cytoskeletal organization, pathways in which ANKEF1 is implicated.
ANKEF1 encodes a protein characterized by ankyrin repeat domains and EF-hand motifs, structural elements that confer protein-protein interaction capabilities and calcium-binding potential. The EF-hand domain suggests regulation by intracellular calcium fluxes, positioning ANKEF1 within calcium-mediated signaling cascades. Predicted interactions with calmodulin or ankyrin family members may link ANKEF1 to calcium/calmodulin-dependent pathways, including cytoskeletal remodeling and signal transduction. Although downstream targets remain uncharacterized, the domain architecture implies a scaffolding function that organizes multiprotein complexes in response to calcium, potentially influencing cellular architecture and signaling dynamics.
In the HEK293T background, knockout of ANKEF1 allows functional dissection of its role in a human epithelial kidney cell model that endogenously expresses relevant signaling components. The HEK293T line??s robust growth and ease of manipulation facilitate downstream analyses, while the renal epithelial origin provides a physiologically pertinent environment for exploring calcium-dependent processes. This polyclonal knockout population avoids biases associated with clonal selection, enabling the study of ANKEF1 in a more heterogeneous, genetically diverse cell pool that better reflects the complexity of tissue-level responses.
These polyclonal knockout cells are suitable for investigating ANKEF1’s role in calcium signaling, cytoskeletal organization, and protein?Cprotein interaction networks. Representative assays include western blotting for protein expression analysis, RT-qPCR for mRNA quantification, immunofluorescence for subcellular localization studies, co-immunoprecipitation for probing interactions with factors such as calmodulin, and live-cell calcium imaging to monitor intracellular calcium dynamics. For additional product details or technical support, please contact Ascent Research.
